Redox control of Nrf2/ARE by thioredoxin and GSH

硫氧还蛋白和 GSH 对 Nrf2/ARE 的氧化还原控制

• 批准号: 6790127
• 负责人: JASON M HANSEN
• 金额: $ 4.73万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6790127
• 关键词: HeLa cells; biological signal transduction; confocal scanning microscopy; gene expression; genetic regulation; glutathione; immunofluorescence technique; intermolecular interaction; oxidation reduction reaction; postdoctoral investigator; protein structure function; thioredoxin; toxicology; transcription factor; transfection /expression vector

DESCRIPTION (provided by applicant): Reactive oxygen species (ROS) have been implicated in teratogenesis. Responsive detoxification systems are paramount for embryonic protection and development. NRF2, a redox-sensitive transcription factor that binds to the ARE to upregulate phase II detoxification enzymes, may rely upon glutathione (GSH) and the oxidoreductase, thioredoxin (TRX1), for signal transduction, but this relationship is not completely defined. I propose to study the role of GSH/GSSG and TRX1 in NRF2 signal transduction. Due to the redox nature of both NRF2 activation and DNA binding, my hypothesis is that GSH/GSSG regulates NRF2 activation, while TRX1 regulates NRF2/DNA binding. I will test this hypothesis by chemical modification of GSH (oxidants, reductants, inhibitors) to change the GSH/GSSG ratios and concentrations to evaluate NRF2 activation and transfection of wild type and active site mutant forms of TRX1 engineered to contain both nuclear localization and nuclear export sequences to evaluate DNA binding. Co-transfection with an ARE4-Luc reporter will determine NRF2 activity. The roles of GSH and cytosolic and nuclear TRX during NRF2 activation and binding will provide an essential foundation for future developmental toxicology.

描述（由申请人提供）：活性氧（ROS）与致畸有关。响应性解毒系统对于胚胎保护和发育至关重要。 NRF2 是一种氧化还原敏感转录因子，与 ARE 结合以上调 II 相解毒酶，可能依赖谷胱甘肽 (GSH) 和氧化还原酶硫氧还蛋白 (TRX1) 进行信号转导，但这种关系尚未完全确定。 我建议研究GSH/GSSG和TRX1在NRF2信号转导中的作用。由于 NRF2 激活和 DNA 结合的氧化还原性质，我的假设是 GSH/GSSG 调节 NRF2 激活，而 TRX1 调节 NRF2/DNA 结合。我将通过化学修饰 GSH（氧化剂、还原剂、抑制剂）来测试这一假设，以改变 GSH/GSSG 比率和浓度，以评估 NRF2 激活以及野生型和 TRX1 活性位点突变形式的转染，这些 TRX1 被设计为包含核定位和核导出序列以评估 DNA 结合。与 ARE4-Luc 报告基因共转染将测定 NRF2 活性。 GSH 以及胞质和核 TRX 在 NRF2 激活和结合过程中的作用将为未来的发育毒理学提供重要基础。